Figure 4. Combination αCTLA-4/α4-1BB therapy enhances intra-tumoral T-cell cytokine production and proliferation.
Mice challenged with 2.5×105 B16-OVA cells and treated on days 6, 9 and 12, were sacrificed on Day 14. TIL were purified from 5–10 pooled tumors per group and enriched using the Miltenyi T-cell purification kit. 2×106 TIL were restimulated with 7.5×105 DC (1∶1 mix of MHC-I and MHC-II peptide pulsed DCs) for 8 hours in the presence of BD GolgiPlug. Cells were fixed using the FoxP3 kit and analyzed by flow cytometry for lymphocyte markers and intracellular IFN-γ and TNF-α production for A) CD8s and B) CD4 Teffs and also Ki67 expression for C) CD8s and D) CD4 Teffs. Data shown is from 4 independent experiments. All means shown are +/− S.E.M.